Photodynamic Eradication of Trichophyton rubrum and Candida albicans.

Conventional methods of onychomycosis treatment are ineffective in some cases because the cure of onychomycosis very often depends on the patient's individual response to the treatment; therefore, there is a crucial need to research and develop new methods of onychomycosis therapy. One of the most innovative treatments is photodynamic therapy (PDT) using photosensitizers (PSs). However, effective treatment depends on the correct choice of photosensitizer and substances that improve the characteristics of the final formulation. The aim of our work was to find an effective formulation for the treatment of onychomycosis. To achieve this goal, we tested the effect of three types of PSs, rose Bengal (RB), malachite green oxalate (MGO), and methylene blue (MB), on Candida albicans. The most effective PS was RB, and so the study was continued with Trichophyton rubrum. Additional comparative studies were carried out on substances included in the formulation (urea and thiourea), focusing on their antifungal activity, which can improve penetration through the nail plate. The composition of the formulation that achieved 100% eradication of Trichophyton rubrum under our conditions consisted of 150 µM RB, 5% urea, and 0.5% thiourea in glycerol/water (70/30%, w/w) solution. A white luminescent lamp was used as a light source (1.9 ± 0.1 mW cm-2). Stability of the formulation was checked. The selected formulation shows potential for future simplification and acceleration of PDT treatment of onychomycosis.

Keratin Biomembranes as a Model for Studying Onychomycosis.

Difficulties in obtaining human nails that are large enough for examining the penetration of drug formulations led us to produce keratin films regenerated from human hair. We assume that these films can simulate human nail plates in drug penetration and permeation tests and can serve as a biological model for studying onychomycosis. The films were formed from keratin extracted from human hair using dithiothreitol, urea and thiourea. The obtained keratin extract was dispensed into Teflon rings and dried at 40 °C and then cured at 110 °C. The structure, surface morphology, chemical characterization and thermal stability of the films were characterized and were compared to those of human nail, hair and bovine hoof samples using SDS-electrophoresis, scanning electron microscopy (SEM), X-ray diffraction analysis (XRD), Fourier transform infrared spectroscopy (FTIR) and thermogravimetric analysis (TGA). The structure of the obtained films was found to be closer to human nails than to hair or bovine hooves. The keratin films were infected with Trichophyton rubrum and were proven to be appropriate for serving as a model for studying onychomycosis.

Bioactive Coating on Ti Alloy with High Osseointegration and Antibacterial Ag Nanoparticles.

Titanium alloys have advanced mechanical properties jointly with high biocompatibility that make them eminently suitable for biomedical applications such as dental and orthopedic implants. Improvement in their osseointegration with human bone can be achieved by the development of hydroxyapatite (HAp) on a Ti alloy surface using different methods of deposition. However, plasma electrolytic oxidation (PEO) treatment has been found to be one of the most promising techniques, due to the formation of high bonding between the bone and the Ti surface. Along with this high bonding, an antibacterial ability of the surface to prevent bacterial infection is also essential. Silver, which is a widely applicable antibacterial agent, was used in this work. First, a titanium oxide coating containing calcium and phosphorus and Ag nanoparticles was formed by PEO treatment. Then, Ti alloy was subjected to hydrothermal treatment to ensure a crystalline formation of HAp. Morphology and phase composition investigations detected the presence of HAp crystals in the coating along with antibacterial agents of silver nanoparticles. The biocompatibility and bioactivity of the created coating were examined by contact angle (CS) measurement and electrochemical impedance spectroscopy (EIS). It was shown that the coating was extensively grown after exposure of the alloy to simulated body fluid (SBF) solution for 7 days with no effect on the Ag nanoparticles. An antibacterial test using Staphylococcus aureus and Escherichia coli revealed that the coating containing Ag nanoparticles has more significant antibacterial effectiveness compared to a coating that does not contain silver.

Polymer-immobilized photosensitizers for continuous eradication of bacteria.

The photosensitizers Rose Bengal (RB) and methylene blue (MB), when immobilized in polystyrene, were found to exhibit high antibacterial activity in a continuous regime. The photosensitizers were immobilized by dissolution in chloroform, together with polystyrene, with further evaporation of the solvent, yielding thin polymeric films. Shallow reservoirs, bottom-covered with these films, were used for constructing continuous-flow photoreactors for the eradication of Gram-positive Staphylococcus aureus, Gram-negative Escherichia coli and wastewater bacteria under illumination with visible white light using a luminescent lamp at a 1.8 mW·cm⁻² fluence rate. The bacterial concentration decreased by two to five orders of magnitude in separate reactors with either immobilized RB or MB, as well as in three reactors connected in series, which contained one of the photosensitizers. Bacterial eradication reached more than five orders of magnitude in two reactors connected in series, where the first reactor contained immobilized RB and the second contained immobilized MB.